Water level or water surface elevation (WSE) is an important state variable of rivers, lakes, and wetlands. Hydrodynamic models of rivers and streams simulate WSE and can benefit from spatially distributed WSE observations, to increase model reliability and predictive skill. This has been partially addressed by satellite radar altimetry, but satellite altimetry is unable to deliver useful data for small rivers. To overcome such limitations, we deployed a radar altimetry system on an unmanned aerial vehicle (UAV), to map spatially distributed WSE. We showed that UAV altimetry can provide observations of WSE with a very high spatial resolution (ca. 0.5 m) and accuracy (ca. 3 cm), in a time-saving and cost-effective way. Furthermore, we investigated the value of this dataset for the calibration and validation of hydrodynamic models. Specifically, we introduced spatially distributed roughness parameters in a hydrodynamic model and estimated these parameters, using the observed WSE profiles along the stream as input. A case study was conducted in the Åmose stream, Denmark. The results showed that UAV-borne WSE can identify significant variations of the Manning–Strickler coefficients, along this small and highly vegetated stream and over time. Moreover, the model performed extremely well using distributed roughness coefficients, but it could not reproduce WSE satisfactorily using uniform roughness. We concluded that distributed roughness coefficients should be considered, especially for small vegetated rivers, to improve model performance, both locally and globally. Spatially distributed parameterizations of the effective channel roughness could be constrained with UAV-borne WSE. This study demonstrated for the first time that UAV-borne WSE can help to understand the variations of hydraulic roughness, and can support efficient river management and maintenance.